Fluorescence Lifetime Macro Imager for Biomedical Applications

Rajannya Sen; Alexander V. Zhdanov; Ciaran Devoy; Mark Tangney; Liisa M. Hirvonen; Andrei Nomerotski; Dmitri B. Papkovsky

doi:10.3791/64321

JoVE Journal
Biology

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JoVE Journal Biology

Fluorescence Lifetime Macro Imager for Biomedical Applications

用于生物医学应用的荧光寿命宏观成像仪

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06:01 min

April 7, 2023

DOI: 10.3791/64321-v

Rajannya Sen¹, Alexander V. Zhdanov¹, Ciaran Devoy², Mark Tangney², Liisa M. Hirvonen³, Andrei Nomerotski⁴, Dmitri B. Papkovsky¹

¹School of Biochemistry and Cell Biology,University College Cork, ²Cancer Research@UCC,University College Cork, ³Centre for Microscopy, Characterisation and Analysis (CMCA),The University of Western Australia, ⁴Physics Department,Brookhaven National Laboratory

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Overview

This study presents a new optical imager designed for macroscopic photoluminescence lifetime imaging, particularly useful in biological samples like live animal tissue. The protocol allows for detailed mapping of phosphorescence lifetime and oxygen concentration, which is crucial for understanding various biological processes.

Key Study Components

Research Area

Biological imaging
Phosphorescence lifetime analysis
Oxygen concentration measurement

Background

Macroscopic imaging is essential for studying complex biological samples.
Existing methods may not efficiently analyze long-decaying samples.
This research aims to enhance visualization techniques in live tissues.

Methods Used

Optical imaging with time-correlated single-photon counting (TCSPC) mode
Application on live animal tissues
Use of custom-designed imaging modules and data processing software

Main Results

Effective visualization of lifetime and oxygen distribution
Ability to generate phosphorescence lifetime images with high contrast
Successful application in observing hypoxic conditions in tissue

Conclusions

The study demonstrates an innovative imaging approach for biological research.
Potential applications for diagnosing and treating tissue hypoxia-related conditions.

Frequently Asked Questions

What is the main application of the optical imager?

It is used for the macroscopic imaging of phosphorescence lifetimes and oxygen concentrations in biological tissues.

How does the imaging module operate?

It operates in TCSPC mode for accurate lifetime distribution visualization.

Can this method be used on other models apart from animal tissues?

Currently, it is limited to animal models, but there is potential for broader applications.

What are the key technological components involved?

Key components include a photomultiplier tube, LEDs, and a compact imaging module.

What biological phenomena can this method help study?

It can aid in the study of conditions like cancer, stroke, and inflammation related to tissue hypoxia.

How is data processed after imaging?

Data is processed using custom software to analyze phosphorescence decay and generate images.

What is the significance of phosphorescence lifetime imaging?

It provides insights into the dynamics of oxygen levels in biological tissues, which is crucial for various research fields.

本文描述了一种新型快速光学成像仪在长衰变发射样品的宏观光致发光寿命成像中的应用。描述了集成、图像采集和分析程序，以及用于成像的传感器材料的制备和表征以及成像仪在研究生物样品中的应用。

我们的协议通过专用探头和传感材料提供宏观物体（特别是活体动物组织和整个动物）中磷光寿命和氧气浓度的详细二维映射。这些信息对于许多研究领域都很重要。我们的协议使用集成且紧凑的成像模块，该模块在TCSPC模式下运行，并提供复杂生物样品（如生命组织）中寿命和氧气分布的简单准确的可视化。

拿起板球适配器并从不同侧面检查它。卸下正面 C 接口适配器以显示安装在板球中的 Photonis PP0360EF增强器，然后将 C 接口适配器放回原处。拆下 Cricket 的正面 C 接口适配器并插入 650 正负 50 纳米发射滤光片。

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